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A method for the intermediate recovery of plasmid DNA (pDNA) from alkaline lysates is described that comprises differential isopropanol precipitation steps. In a first low-cut precipitation, a smaller amount of isopropanol (20% v/v) is used so that only high molecular weight RNA precipitates. After solid liquid separation, a high-cut precipitation is performed by bringing isopropanol concentration to 70% v/v to precipitate pDNA. Tests made with lysates show that the differential precipitation increases purity threefold compared to the conventional one-step precipitation at 70% v/v without affecting pDNA recovery (>80%).


Alexandra Wagner, A Rita Silva-Santos, Sara Sousa Rosa, Sophie Gierak, Ana M Azevedo, Duarte Miguel F Prazeres. Primary Purification of Plasmid DNA Using Differential Isopropanol Precipitation. Methods in molecular biology (Clifton, N.J.). 2021;2197:151-165

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PMID: 32827136

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